Cytotoxic and Apoptotic Effect of Nanoclinoptilolite on Canine Osteosarcoma Cell Lines.

INTRODUCTION: Clinoptilolite has antiviral, antibacterial, anti-inflammatory, antidiabetic, and anticancer properties due to its biological activities. In various cancer cell culture studies, it has been reported effective against tumour cells and gave positive results in treatment of various tumours in dogs. No study was found on the effects of the nanoparticulate form, nanoclinoptilolite, on cancer cells. The aim of this study was to determine its cytotoxic and apoptotic effects in canine osteosarcoma (OSA) cell culture. MATERIAL AND METHODS: Doses at 50% inhibitory concentration were determined by measuring the dose- and duration-dependent cytotoxicity of nanoclinoptilolite on canine D-17 osteosarcoma cells by methylthiazol tetrazolium (MTT) test for 24 h, 48 h, and 72 h. Murine caspase-3 and -7 activity and expression levels of the BAX and BCL2 genes were measured using RT-PCR to investigate the apoptotic effect. RESULTS: Nanoclinoptilolite decreased cell viability and induced caspase-3- and -7-mediated apoptosis in treated canine OSA cells. Furthermore, its application to canine OSA cells downregulated the expression of BCL2 and upregulated the expression of proapoptotic BAX. CONCLUSION: Clinoptilolite, which was previously demonstrated to have anticancer properties, decreased cell viability effectively and rapidly and increased the apoptotic cell ratio in a novel use in nanoparticle form, exhibiting this effect by increasing the BAX/BCL2 ratio.

The protective effects of vitamin C on the DNA damage, antioxidant defenses and aorta histopathology in chronic hyperhomocysteinemia induced rats.

The aim of this study was to investigate the protective effect of vitamin C towards hyperhomocysteinemia (hHcy) induced oxidative DNA damage using the comet assay. The increase in plasma homocysteine levels is an important risk factor for vascular and cardiovascular diseases through free radical production. This study was also conducted to investigate the histopathological changes in the thoracic aorta and the oxidant/antioxidant status in heart, liver and kidney tissues. Twenty-four adult male Wistar rats were divided as control, hHcy and hHcy+vitamin C group. Chronic hHcy was induced by oral administration of l-methionine (1g/kg/day) for 28 days. Vitamin C was given 150mg/kg/day within the specified days. DNA damage was measured by use of the comet assay in lymphocytes. Levels of malondialdehyde (MDA) and glutathione (GSH) as well as catalase (CAT) and superoxide dismutase (SOD) activities were determined in heart, liver and renal tissues. Results show that l-methionine administration significantly increased % Tail DNA and Mean Tail Moment in hHcy group as compared with other groups. Vitamin C treatment significantly decreased the high MDA levels and increased activity of antioxidant enzymes in tissues. Aortic diameter and thickness of aortic elastic laminae were significantly lower in hHcy+vitamin C group. Comet assay can be used for the assessment of primary DNA damage caused by hHcy. Histopathological findings showed that vitamin C may have a preventive effect in alleviating the negative effects of hHcy. Vitamin C might be useful in the prevention of endothelial dysfunction caused by hHcy.

Acute phase protein levels in pregnancy and oestrus cycle in bitches.

The purpose of this study was to evaluate the acute phase protein (serum amyloid A, C-reactive protein, haptoglobin, ceruloplasmin and fibrinogen) profiles in pregnant dogs as well as bitches at various stages of the estrus cycle and to verify possible relationship(s) among the APPs profile and hormonal status. A total of 60 bitches of various breeds were enrolled. The animals were divided into six groups. B-mode ultrasonographic scanner (concept M/C) was used for pregnancy diagnosis, involving the first and second halves of pregnancy. Proestrus, estrus, diestrus and anestrus stages of sexual cycles were differentiated by use of vaginal cytology and blood progesterone and estradiol 17-beta levels. Serum C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin, ceruloplasmin and fibrinogen levels were determined. No statistically significant differences were observed between the groups with regard to the serum SAA and CRP concentrations. Haptoglobin, ceruloplasmin and fibrinogen concentrations were elevated in pregnant dogs compared to non-pregnant groups', independently from the pregnancy stage. No statistically significant differences in acute phase protein concentrations in dogs at various stages of estrus could be detected. Results of this study indicated that ceruloplasmin and fibrinogen levels may have potential importance in the diagnosis of pregnancy, and acute phase proteins may be used for monitoring the health status of the pregnant dogs.

Effects of xylene and formaldehyde inhalations on oxidative stress in adult and developing rats livers.

In this study, it was aimed to demonstrate the possible oxidative stress caused by exposure of xylene and formaldehyde (HCHO) on liver tissue, and on body and liver weights in adult as well as developing rats. The rats (96 female Sprague-Dawley) were randomly divided into four groups: embryonic day 1 (Group 1), 1-day-old infantile rats (Group 2), 4-week-old rats (Group 3) and adult rats (Group 4). The animals were exposed to gases of technical xylene (300 ppm), HCHO (6 ppm) or technical xylene + HCHO (150 ppm + 3 ppm), 8 hours per day for 6 weeks. Superoxide dismutase (SOD) and catalase (CAT) activities, and glutathione (GSH) and malondialdehyde (MDA) levels were evaluated. In addition, body and liver weights were determinated. Compared to the control animals, body and liver weights were decreased in the embryonic day 1 group (P < 0.001, P < 0.01, respectively) and the 1-day-old infantile group (P < 0.001). Liver weight was increased in the 4-week-old group (P < 0.01). SOD activities were decreased in the 4-week-old rats exposed to HCHO (P < 0.01). CAT activities increased in the embryonic day 1 group (P < 0.05). GSH levels were decreased in the 1-day-old infantile group (P < 0.01), and MDA levels was increased in the embryonic day 1 group (P < 0.05) as compared with the respective control groups. As to GSH and MDA levels in adult and 4-week-old animals, no statistically significant differences were observed (P > 0.05). The present study indicates that exposures to xylene, HCHO and a mixture of them are toxic to liver tissue, and developing female rats are especially more adversely affected. Furthermore, the results of this study show that adult female rats could better tolerate the adverse effects of these toxic gases.

Effects of xylene and formaldehyde inhalations on renal oxidative stress and some serum biochemical parameters in rats.

In this study, it was aimed to demonstrate the possible renal oxidative stress and some serum biochemical parameters and their alterations caused by the exposure to xylene and formaldehyde (HCHO) in rats. Weighing 150-200g, 12-week-old, 24 female Sprague-Dawley rats were used. The rats were randomly divided into four groups: Group 1 (control), Group 2 (300-ppm technical xylene), Group 3 (6-ppm HCHO) and Group 4 (150-ppm technical xylene + 3-ppm HCHO). The animals were exposed to gases eight hours per day for six weeks. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities, glutathione (GSH) and malondialdehyde (MDA) levels were measured. In addition, serum total protein, albumin, urea and creatinine levels were evaluated. Compared with the control animals, urea levels increased significantly in all groups (P < 0.001). GSH activities and MDA levels increased in xylene and xylene + HCHO groups (P < 0.05). No statistically considerable differences were found in SOD, CAT and GSH-Px activities, total protein, albumin and creatinine levels among all groups (P > 0.05). The present study indicates but not statistically confirms the renal toxicity of the exposures to xylene, HCHO and a mixture of them.

Dogs with Hepatozoon canis respond to the oxidative stress by increased production of glutathione and nitric oxide.

Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon spp. It has been reported in the United States, southern Europe, the Middle East, Africa and the Far East. In Turkey, canine hepatozoonosis was reported for the first time in 1933. In the present study, serum glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO) and ceruloplasmin levels were analysed in 14 dogs infected with Hepatozoon canis as well as in 10 healthy dogs. Blood smears were prepared from peripheral blood and ticks were collected for identification in the laboratory. Rhipicephalus sanguineus was found only on diseased dogs. No ticks were observed on healthy dogs. The diagnosis of H. canis is made mainly by the detection of gametocytes within neutrophils and monocytes. The haematological diagnosis was confirmed using PCR analyses by amplifying a partial 18S rRNA gene sequence of Hepatozoon spp. Infection was detected in 14 animals. Compared to controls, the serum GSH, MDA and NO levels in infected animals increased significantly (p<0.05, <0.01 for MDA), whereas the concentrations of ceruloplasmin in diseased animals remained unaltered. The results of the present study suggest that in dogs infected with H. canis increased levels of GSH, MDA and NO may be related to host's defences against parasitic infection.